KR20090013664A - Novel indoledione derivative compound and pharmaceutical composition comprising the same for the prevention and treatment of vascular smooth muscle cell hyper-proliferative disease - Google Patents

Abstract

A novel indoledione derivative compound is provided to improve the anti-proliferative activity against the vascular smooth muscle cell, thereby preventing and treating the hyper-proliferative disease of the vascular smooth muscle cell and secondly generated various cardiovascular diseases therefrom. The indoledione derivative compound having improved anti-proliferative activity is represented by the chemical formula(I), wherein X and Y are each independently C or N; when X is C and Y is N, R1 is H or C1-C4 linear or branched alkyl group, and R2 is H, C1-C4 linear or branched alkyl group or alkoxy group; and when X is N and Y is C, R1 is H or C1-C4 linear or branched alkyl group, and R2 is H, halogen, or C1-C4 linear or branched alkyl group.

Description

Novel Indoledione Derivative Compound and Pharmaceutical Composition Comprising the Same for the Prevention and Treatment of Vascular Smooth Muscle Cell Hyper-Proliferative Novel Indoledione Derivative Compound and Pharmaceutical Composition Comprising the Same for the Prevention and Treatment of Vascular Smooth Muscle Cell Hyper-Proliferative Disease}

The present invention provides a novel indole dione derivative compound, a composition for inhibiting vascular smooth muscle cell proliferation comprising the compound as an active ingredient, and a pharmaceutical composition for preventing or treating vascular smooth muscle cell abnormal proliferative disease containing the compound as an active ingredient. It is about.

Cardiovascular diseases, including heart failure, hypertensive heart disease, arrhythmia, congenital heart disease, myocardial infarction and angina, and vascular diseases such as arteriosclerosis, stroke, and peripheral vascular disease, are widely referred to as ischemic cardiovascular disease. . These diseases occur in various age groups and can lead to death with serious sequelae if not treated in a timely manner. Due to westernized dietary and lifestyle changes, the incidence and mortality rate of cardiovascular diseases continues to increase rapidly. However, effective therapeutics for treating such diseases have not been developed.

Current treatments for coronary artery disease include pharmacotherapy, gene therapy, percutaneous transluminal coronary angioplasty and stenting (PTCA), and reperfusion therapy of CABG (coronary artery bypass graft). revascularization therapy is underway. However, even after reperfusion therapy, the drug should be continued and there is a possibility of recurrence of coronary artery disease.

In particular, percutaneous coronary angioplasty (PTCA) has been widely used since 1977 as a treatment for patients with coronary atherosclerosis (Gurentzig, Lancet, 4, 263, 1978). After 3-6 months, restenosis occurs in about 40% of patients (Ryan et al., J. Am . Coll . Cardiol . , 22. 2033-2054, 1993). The mechanism of the development of restenosis after PTCA is that the cytokine is produced and secreted from various vascular endothelial cells, activated platelets or macrophages damaged by a balloon, which promotes the migration and proliferation of vascular smooth muscle cells. It is known that the production and secretion of extracellular matrix (ECM) is promoted, leading to thickening of the vascular lining and developing into restenosis (Godfried et al, Am . Heart J., 129, 203-210, 1995).

Normal vascular smooth muscle cells do not proliferate, but endothelial cell membranes are damaged through stents, resulting in multiple stages of signal transduction resulting in the division, migration and proliferation of vascular smooth muscle cells. The mechanisms of vascular smooth muscle cell proliferation include the removal of proliferation inhibitory factors and the activation of proliferation-inducing factors due to normal endothelial cell damage, the transmission of proliferation-induced signals through receptors on the surface of vascular smooth muscle cells, and the proliferation delivered into the nucleus of vascular smooth muscle cells. And a change in the cell cycle caused by the trigger signal. Normal endothelial cells secrete substances that inhibit the proliferation of vascular smooth muscle cells.When the endothelial cells are damaged, their secretion is suppressed and platelet derived growth factor, which is released from activated platelets, is contained in plasma. Various cytokines (cytokine), etc. are known to induce proliferation of vascular smooth muscle cells.

To date, attempts have been made to prevent restenosis using antiplatelet agents, anticoagulants, cholesterol biosynthesis inhibitors, and antiallergic agents (Lefkovits et al., Progr . Cardiovas . Dis ., 40, 141-158, 1997). There is research on the prevention of restenosis of natural oil-derived components such as fish oil, vitamin C and vitamin E. However, these drugs have not yet obtained sufficient results in clinical practice.

Taxol (Herdeg et al., Zeischrift) fur Kardiologies , 89, 390-397, 1999), heparin, EPA, estrogens, etc., are under research and development as inhibitors of vascular restenosis, but these have not been shown to be effective. However, Korean Patent Registration No. 478671 discloses a pharmaceutical composition for preventing and treating vascular restenosis including clotrimazole as an active ingredient, and Korean Patent Registration No. 516026 discloses 3-deoxyadenosine. A composition for preventing and treating vascular restenosis as an active ingredient is disclosed, and Korean Unexamined Patent Publication No. 2001-110793 discloses an agent for preventing and treating vascular stenosis as an active ingredient of a compound having Rho kinase inhibitory activity. In addition, Korean Patent Laid-Open Publication No. 2003-46314 discloses the prevention and treatment of vascular dysplasia such as transplanted vascular disease, restenosis, in-stent restenosis, and pulmonary hypertension with antithrombin as an active ingredient. A composition for a treatment is disclosed, and Korean Patent Laid-Open Publication No. 2005-23249 discloses prevention of vascular disease such as stenosis and / or redisclusion after percutaneous coronary angioplasty using an endovascular stent containing a retinoid and a retinoid action modifier. And / or a pharmaceutical composition for the treatment is disclosed, Korean Patent Publication No. 2005-43183 discloses a pharmaceutical composition for preventing and treating vascular restenosis, including curcumin (curcumin). However, in addition to preventing vascular restenosis, these vascular restenosis inhibitors have various side effects such as inhibiting wound regeneration, vascular injury, hepatotoxicity, kidney damage, and increased bleeding by platelet aggregation. Although research is being actively conducted to develop a substance capable of inhibiting vascular restenosis derived from the various proven natural products, no research results have been reported. Therefore, there is a constant need to develop materials that can safely and effectively inhibit vascular restenosis.

In addition, atherosclerosis is a disease in which the elasticity of the artery wall is lost and the wall is hardened and hardened. The arteriosclerosis exhibits various symptoms depending on the organs involved, but is most commonly caused by ischemia due to a lack of blood supply and loss of elasticity. Because of the symptoms of rupture hemorrhage. In particular, if the coronary artery causes arteriosclerosis and blood flow is reduced, angina pectoris is felt during exercise, and if it is further reduced, it becomes unstable angina, which causes pain even at rest, and if very blocked, myocardial infarction This can put your life at risk.

In the meantime, researches on treating such arteriosclerosis have been conducted. In the 1970s, the tube was inserted into the heart, angiography was discovered, and the lesion was found. The idea of a method of physically widening the balloon was introduced. Recently, drug-coated stents have been used to increase the success rate of the procedure, and the success rate of the procedure in a wider range of situations is increasing, leading to the treatment of heart disease. Although the surgical technique for atherosclerosis has made such a lot of progress, the academic achievements to prevent and control arteriosclerosis itself still have much to be solved, and studies on the causes and treatment of atherosclerosis, Research and application of effective antithrombotic and anticoagulant agents to inhibit thrombus formation are needed.

Throughout this specification, many papers and patent documents are referenced and their citations are indicated. The disclosures of cited papers and patent documents are incorporated herein by reference in their entirety, and the level of the technical field to which the present invention belongs and the contents of the present invention are more clearly explained.

The present inventors have made diligent efforts to develop a substance that can effectively treat and prevent diseases caused by excessive proliferation of vascular smooth muscle cells. As a result, the indoldion derivative compounds synthesized by the present inventors have been described as vascular smooth muscle cells (VSMC). The present invention was completed by experimentally confirming that there is a growth inhibitory activity of.

Accordingly, an object of the present invention is to provide a novel indole dione derivative compound.

Another object of the present invention to provide a composition for inhibiting vascular smooth muscle cell proliferation comprising a novel indole dione derivative compound as an active ingredient.

Another object of the present invention to provide a pharmaceutical composition for the prevention and treatment of vascular smooth muscle cell abnormal proliferative disease containing a novel indole dione derivative compound as an active ingredient.

The objects and advantages of the invention will become apparent from the following detailed description, claims and drawings.

According to one aspect of the present invention, the present invention provides a novel indoldione derivative compound represented by the following formula (I).

[Formula I]

However, in Formula I, X and Y are each independently C or N, and when X is C and Y is N, R ₁ Is H or a C ₁ -C ₄ straight or branched alkyl group, R ₂ Is H, a C ₁ -C ₄ straight or branched alkyl group or an alkoxy group; When X is N and Y is C, R ₁ is H or a C ₁ -C ₄ straight or branched alkyl group, and R ₂ is H, halogen or a C ₁ -C ₄ straight or branched alkyl group.

The present inventors have made diligent efforts to develop substances capable of effectively inhibiting diseases caused by excessive proliferation of vascular smooth muscle cells (VSMCs). As a result, the inventors have synthesized superior vascular smooth muscle. The present invention was completed by experimentally confirming that the cell proliferation inhibitory activity is present.

The indoldione derivative compounds of the formula I are novel compounds synthesized by the inventors for the first time.

Among the indoldione derivative compounds of the above formula (I) of the present invention, when X is C and Y is N, the compound is represented by the following formula (Ia).

[Formula Ia]

The compound of Formula Ia may be synthesized by the following Scheme 1.

Scheme 1

In the above scheme A is a C _{1 - 4} alkyl-dicyano acetate / ethanol or cyano acetate / ethanol, b is C _{1 -} is _{4-alkoxyphenyl-amine} / ethanol or phenylamine / ethanol _{- 4} alkylphenyl amine / ethanol, C _1.

Compound 1 (6,7-dichloro-5,8-quinolinone) used as starting material in the above reaction scheme is known in the art ( J. Am , Chem . Soc , 82, 1155 (1960). )] And [ J. Med . Chem . 1986 , 29, 1329]).

In the above scheme 2, the compound is the compound C 1 ₁ in an alcohol _solvent may be prepared by reaction with _{4-alkyl-dicyano} acetate. The C _{1 -4} alkyl is straight chain or branched chain alkyl. If desired, the reaction can be carried out in the presence of dimethylsulfoxide (DMSO) or glacial acetic acid. As the alcohol solvent, methanol, ethanol, isopropanol and mixtures thereof can be used, preferably ethanol.

In the above scheme, Compound 3 Compound 2 may be prepared by C _{1 -4} alkyl phenyl amine, phenyl-C _{1 -4} alkoxy or phenyl amines and amine react in an alcohol solvent. The C _{1 -4} alkyl or C _{1 -4} alkoxy is a straight or branched C _{1 -4} alkyl or alkoxy. As the alcohol solvent, methanol, ethanol, isopropanol and mixtures thereof can be used, preferably ethanol. The reaction can be carried out for 2-6 hours in the temperature range of room temperature to 100 ℃, it is preferable to react for 3-5 hours while refluxing at the boiling point of the solvent used. The C _{1 -4} alkyl phenyl amine can be used in an amount of 1 to 1.2 equivalents, it is 1.1 to 1.5 equivalents of a compound represented by the vertical wind 2.

Preferred compounds of the invention are those wherein R ₁ Is H or -CH ₃ , R ₂ H, -CH ₃ Or a compound represented by -OCH ₃ , and more preferably a compound represented by the following general formula (II), (III), or (IV).

[Formula II] [Formula III]

[Formula IV]

Among the indoldione derivative compounds of the general formula (I) of the present invention, compounds in which X is N and Y is C are represented by the following general formula (Ib).

[Formula Ib]

The compound of Formula Ib (1H-pyrrolo [3,2-g] isoquinoline-4,9-diones derivative) may be prepared by the following Scheme 2.

Scheme 2

In the above scheme A is a C _{1 - 4} alkyl-dicyano acetate / ethanol or a cyano quinoa three Tate / ethanol, b is C _{1 - 4} alkyl, phenyl amine / ethanol, halogenated phenylamine / ethanol or phenylamine / ethanol.

6,7-dichloroisoquinoline-5,8-dione (Compound 4 ), which is used as a starting material in the preparation method according to the present invention, is known in the art. As a compound, it may be conveniently prepared from 5-hydroxyisoquinoline. The method for preparing the 6,7-dichloroisoquinoline-5,8-dione compound can be synthesized with reference to J. Med . Chem . 1986 , 29, 1329.

Referring to the preparation method of the compound of the present invention according to the above scheme in more detail as follows. The compound represented by the compound 5 is 6,7-dichloroisoquinoline-5,8-dione (6,7-dichloroisoquinoline-5,8-dione) and ethyl-cyanoacetate (ethyl) represented by the compound 4 cyanoacetate) or methyl cyanoacetate is prepared by reaction in the presence of an alcohol solvent or dimethylsulfoxide (DMSO) or glacial acetic acid. In this case, as the alcohol solvent, lower alcohols having 1 to 3 carbon atoms such as methanol, ethanol, isopropanol, and the like may be used, and ethanol is particularly preferable. The reaction is carried out for 2-6 hours in the temperature range of room temperature to 100 ℃, it is preferable to react for 3-5 hours while refluxing at the boiling point of the solvent used.

In the above reaction scheme 6 compounds may be prepared by the compound 5-C _{1 -4} alkyl phenyl amine, C _{1 -4} halogenated phenyl or phenyl amines and amine react in an alcohol solvent. The C _1-4 alkyl is straight or branched C _1-4 alkyl. As the alcohol solvent, methanol, ethanol, isopropanol and mixtures thereof can be used, preferably ethanol. The reaction can be carried out for 2-6 hours in the temperature range of room temperature to 100 ℃, it is preferable to react for 3-5 hours while refluxing at the boiling point of the solvent used. The C _1-4 alkylphenylamine, C _1-4 halogenatedphenylamine or phenylamine may be used in an amount of 1-1.2 equivalents, preferably 1.1-1.5 equivalents, relative to compound 5 .

Preferred compounds of the invention are those wherein R ₁ Is H or -CH ₃ , R ₂ Is -CH ₃ , It is a compound which is fluorine (F), chlorine (Cl), bromine (Br), or iodine (I), More preferably, it is a compound represented by following formula (V) or (VI).

[Formula Ⅴ] [Formula VI]

According to another aspect of the present invention, the present invention provides a composition for inhibiting vascular smooth muscle cell proliferation comprising an indoldion derivative compound represented by the following formula (I) as an active ingredient.

[Formula I]

However, in Formula I, X and Y are each independently C or N, and when X is C and Y is N, R ₁ Is H or a C ₁ -C ₄ straight or branched alkyl group, R ₂ Is H, a C ₁ -C ₄ straight or branched alkyl group or an alkoxy group; When X is N and Y is C, R ₁ is H or a C ₁ -C ₄ straight or branched alkyl group, and R ₂ is H, halogen or a C ₁ -C ₄ straight or branched alkyl group.

Preferred compounds used as an active ingredient of the composition for inhibiting vascular smooth muscle cell proliferation of the present invention, when X is C and Y is N in Formula I, R ₁ is H or -CH ₃ , R ₂ is a compound represented by H, -CH ₃ or -OCH ₃ , more preferably a compound represented by the formula (II), (III) or (IV).

In addition, another preferred compound used as an active ingredient of the composition for inhibiting vascular smooth muscle cell proliferation of the present invention is R ₁ when X is N and Y is C in the formula (I). Is H or -CH ₃ , R ₂ Is -CH ₃ , It is a compound which is fluorine (F), chlorine (Cl), bromine (Br), or iodine (I), More preferably, it is a compound represented by the said Formula (V) or (VI).

Since the indoldion derivative compound of the present invention has a proliferation inhibitory activity of vascular smooth muscle cells as described in detail below, it can be very useful in various fields in which excessive proliferation of vascular smooth muscle cells needs to be suppressed.

According to another aspect of the present invention, the present invention provides a pharmaceutical composition comprising (a) a pharmaceutically effective amount of an indoldione derivative compound represented by the following formula (I); And (b) provides a pharmaceutical composition for the prevention or treatment of vascular smooth muscle cell hyper-proliferative disorder comprising a pharmaceutically acceptable carrier.

 [Formula I]

However, in Formula I, X and Y are each independently C or N, and when X is C and Y is N, R ₁ Is H or a C ₁ -C ₄ straight or branched alkyl group, R ₂ Is H, a C ₁ -C ₄ straight or branched alkyl group or an alkoxy group; When X is N and Y is C, R ₁ is H or a C ₁ -C ₄ straight or branched alkyl group, R ₂ Is an alkyl group of H, halogen or C ₁ -C ₄ straight or branched chain.

Preferably, the compound used as an active ingredient of the pharmaceutical composition of the present invention, in the above formula (I) when X is C and Y is N, R ₁ is H or -CH ₃ and R ₂ is H, -CH ₃ or- OCH ₃ or a compound, more preferably a compound represented by the general formula ⅱ, ⅲ formula, or formula ⅳ.

In addition, preferably the compound used as an active ingredient of the pharmaceutical composition of the present invention is R ₁ when X is N and Y is C in the formula (I) Is H or -CH ₃ , R ₂ Is -CH ₃ , It is a compound which is fluorine (F), chlorine (Cl), bromine (Br), or iodine (I), More preferably, it is a compound represented by the said Formula (V) or (VI).

Indoldion derivative compounds of the present invention have a proliferation inhibitory activity of vascular smooth muscle cells. According to a specific embodiment of the present invention, the compound of the present invention has an effect of inhibiting the proliferation of aortic smooth muscle cells of the rat induced by platelet-dereived growth factor (PDGF-BB).

According to another specific embodiment of the present invention, the compound of the present invention inhibits the incorporation of [ ³ H] thymidine into DNA during PDGF-BB induced rat aortic smooth muscle cell proliferation .

In the context of the present invention, the term smooth muscle refers to organs such as tubular organs, bladder, abdominal cavity, uterus, reproductive tract, gastrointestinal tract, respiratory tract, vasculature, skin and ciliary muscle, eye iris, etc. Non-striated muscle found in the wall of the patient. The smooth muscle is spindle shaped and is capable of contraction and relaxation.

Smooth muscle cells, which constitute the smooth muscle, are mononuclear cells, and are arranged in the form of sheets or bundles and are connected by gap junctions.

The term vascular smooth muscle cell (VSMC) in the context of the present invention refers to the cells that make up the smooth muscle of the vessel wall.

As used herein, the term VSMC hyper-proliferative disorder refers to a disease or condition caused by excessive proliferation of vascular smooth muscle cells.

Vascular smooth muscle cell aberrant proliferative disease in the present invention includes various diseases. For example, vascular restenosis, vascular stenosis, arteriosclerosis, atherosclerosis, which are directly caused by abnormal proliferation of vascular smooth muscle cells, as well as cardiovascular diseases secondary to stenosis or atherosclerosis, heart failure, myocardium Infarctions, angina pectoris, arrhythmia, hypertensive heart disease, congenital heart disease, stroke, peripheral vascular stenosis, and the like.

Preferably, the vascular smooth muscle cell aberrant proliferative disease treated or prevented by the pharmaceutical composition of the present invention is atherosclerosis, atherosclerosis, vascular restenosis or vascular stenosis.

Atherosclerosis is a disease in which fatty substances are deposited or fibrosis in the inner layers of arteries. On the other hand, restenosis of blood vessels is a disease in which the vascular pathway is narrowed after traumatization. Vascular restenosis occurring after implantation of arteriosclerosis progression and stent is known that due to the secretion of proliferation, migration, and extracellular matrix (extracellular matrix) of vascular smooth muscle cells (Circulation, 1997, 95, 1998-2002 ; J. Clin . Invest . 1997, 99, 2814-2816; Cardiovasc. Res. 2002, 54, 499-502). In order to prevent the progression of arteriosclerosis and vascular restenosis, studies on drugs that inhibit the proliferation of vascular smooth muscle cells have been widely conducted. Currently, several drugs have been used for the treatment of patients (J. Am. Coll. Cardiol., 2002, 39, 183-193). Therefore, it can be seen that the indoldion derivative compounds of the present invention which inhibit the proliferation of vascular smooth muscle cells very efficiently are very effective in the treatment of vascular smooth muscle cell abnormal proliferative diseases.

The pharmaceutical composition for preventing or treating vascular smooth muscle cell abnormality proliferative disease of the present invention comprises an active ingredient and a pharmaceutically acceptable carrier.

Pharmaceutically acceptable carriers included in the pharmaceutical compositions of the present invention are those commonly used in the preparation, such as lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, Calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like It doesn't happen. The pharmaceutical composition of the present invention may further include lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives and the like in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

Suitable dosages of the pharmaceutical compositions of the present invention may vary depending on factors such as the formulation method, mode of administration, age, weight, sex, morbidity, condition of food, time of administration, route of administration, rate of excretion and response to response of the patient. Can be. On the other hand, the oral dosage of the pharmaceutical composition of the present invention is preferably 0.001-100 mg / kg (body weight) per day.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and when administered parenterally, may be administered by intravenous infusion, subcutaneous infusion, intramuscular injection, intraperitoneal injection, transdermal administration, or the like. It is preferable that the route of administration is determined according to the type of the disease to which the pharmaceutical composition of the present invention is applied.

The concentration of the indoldion derivative compound, which is an active ingredient included in the composition of the present invention, may be determined in consideration of the purpose of treatment, the condition of the patient, the period of time, etc., and is not limited to a specific range of concentration. Preferably, however, the indoldione derivative compounds of the present invention may be added at a concentration of 0.5-5 μM in the composition. It is difficult to expect the desired effect when included in a concentration of less than 0.5 μM, it is difficult to expect an increased effect proportional to the increase in concentration when contained in a concentration of more than 5 μM and also increase the manufacturing cost of the product according to the present invention There are disadvantages.

The pharmaceutical compositions of the present invention may be prepared in unit dosage form by formulating with a pharmaceutically acceptable carrier and / or excipient according to methods which can be easily carried out by those skilled in the art. Or it may be prepared by incorporation into a multi-dose container. In this case, the formulation may be in the form of a solution, suspension or emulsion in an oil or an aqueous medium, or may be in the form of extracts, powders, granules, tablets or capsules, and may further include a dispersant or stabilizer.

The present invention provides a novel indole dione derivative compound, a composition for inhibiting vascular smooth muscle cell proliferation comprising the compound as an active ingredient, and a pharmaceutical composition for preventing and treating vascular smooth muscle cell abnormal proliferative disease comprising the compound as an active ingredient. to provide.

Since the derivative compounds of the novel indoldion of the present invention have excellent vascular smooth muscle cell proliferation inhibitory activity, arteriosclerosis, atherosclerosis, vascular restenosis, vascular stenosis, and secondary occurrence due to vascular smooth muscle cell abnormality proliferative diseases It can be very useful for the prevention or treatment of various cardiovascular diseases.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. .

Example  One : methyl  2- (7- chloro -5,8- dihydro -5,8- dioxoquinolin -6- yl ) -2-cyanoacetate synthesis

A is methyl cyanoaccetate / ethanol in the above scheme.

6,7-dichloroquinoline-5,8-dione 590 mg (2.66 mmole) was added to 100 ml abs. After dissolving in ethanol, 0.26358 ml (2.66 mmol) of methyl cyanoaccetate was added thereto. 2 ml of ammonia water was added and reacted for 10 minutes, followed by acidification with dil-HCl. Extraction with MC and separation with EA were performed by column chromatography. The resulting compound was brown oil (32%) in the form, NMR analysis of the compound was as follows.

¹ H-NMR (CDCl _{3 ,} 400 MHz) δ 3.91 (s, 3H, methoxy), 5.50 (s, 1H, CH), 7.80 (q, 1H, 2-pyridine), 8.53 (q, 1H, 2-pyridine), 9.14 (q, 1H, 2-pyridine); MS (m / z) 290 (M ⁺ ).

The synthesized compound was identified as methyl 2- (7-chloro-5,8-dihydro-5,8-dioxoquinolin-6-yl) -2-cyanoacetate.

Example  2 : methyl  2- amino -4,9- dihydro -4,9- dioxo -1-p- tolyl -1H- pyrrolo  Synthesis of [3,2-g] quinoline-3-carboxylate

B in the above scheme is 4-methyl-phenylamine / ethanol.

150 mg of the compound methyl-2- (7-chloro-5,8-dihydro-5,8-dioxoquinolin-6-yl) -2-cyanoacetate synthesized in Example 1 was dissolved in 100 ml abs. Ethanol and stirred Then, 4-methyl-phenylamine corresponding to the equivalent was added thereto and reacted at room temperature for 5 hours. Termination of the reaction was confirmed using TCL. The reaction solution was cooled and concentrated, and then separated by chromatography using chloroform, and then recrystallized with ethanol.

The structure of the synthesized compound was analyzed by NMR. The NMR analysis result was as follows.

mp 262-263 ° C ¹ H-NMR (CDCl _3, 400 MHz) δ 2.37 (s, 3H, methyl), 3.92 (s, 3H, methoxy), 6.98 (d, 1H, benzene), 7.14 (d, 1H, benzene), 7.50 (m, 1H, benzene), 7.66 (q, 1H, 2-pyridine), 7.91 (m, 1H, benzene), 8.43 (q, 1H, 2-pyridine), 9.09 (q, 1H, 2-pyridine); MS (m / z) 361 (M ⁺ ).

Compound synthesized by NMR analysis was identified as methyl-2-amino-4,9-dihydro-4,9-dioxo-1-p-tolyl-1H-pyrrolo [3,2-g] quinoline-3-carboxylate The synthesized compound was named "YSK2821".

Example 3 Methyl 2-amino-4,9-dihydro-1- (4-methoxyphenyl) -4,9-dioxo-1H-pyrrolo [3,2-g] quinoline -3- carboxylate Synthesis of

B in the above scheme is 4-methoxyphenylamine / ethanol.

Methyl 2- (7-chloro-5,8-dihydro-5,8-dioxoquinolin-6-yl) -2-cyanoacetate synthesized in Example 1 150 mg 100 ml abs. After dissolving in EtOH and stirring, 4-methoxyphenylamine corresponding to the equivalent was added thereto and reacted at room temperature for 4-5 hours. Termination of the reaction was confirmed by TCL. The reaction solution was cooled and concentrated, and then separated by column chromatography with EA: HX = 1: 1 solvent and recrystallized with 95% EtOH. The structure of the synthesized compound was analyzed by NMR.

The NMR analysis result was as follows. Dark violet powder (10%); mp 261.1- 261.3 ° C; ¹ H-NMR (CDCl _{3 ,} 400 MHz) δ 3.85 (s, 3H, methoxy), 3.89 (s, 3H, methoxy), 6.99 (m, 1H, benzene), 7.06 (d, 1H, benzene), 7.21 (d, 1H, benzene), 7.27 (m, 1H, benzene), 7.66 (q, 1H, 2-pyridine), 7.96 (s, 2H, NH 2), 8.45 (q, 1H, 2-pyridine), 9.08 (q, 1H, 2-pyridine); MS (m / z) 377 (M ⁺ ).

Compounds synthesized based on the results of NMR analysis were Methyl-2-amino-4,9-dihydro-1- (4-methoxyphenyl) -4,9-dioxo-1H-pyrrolo [3,2-g] quinoline-3- It was identified as carboxylate and named "YSK2822".

Example  4 : Methyl  2- amino -4,9- dihydro -4,9- dioxo -One- phenyl -1H-pyrrolo [3,2-g] quinoline-3- carboxylate Synthesis of

B is aniline / ethanol in the above scheme.

150 mg of Methyl 2- (7-chloro-5,8-dihydro-5,8-dioxoquinolin-6-yl) -2-cyanoacetate synthesized in Example 2 was added to 100 ml abs. After dissolving in EtOH and stirring, an aniline corresponding to the equivalent was added and reacted at room temperature for 4-5 hours. The completion of the reaction was confirmed using TCL. The reaction solution was cooled and concentrated, separated by column chromatography with EA: HX = 1: 1 solvent, and then recrystallized with 95% EtOH.

The structure of the synthesized compound was analyzed by NMR. The NMR analysis result was as follows.

Bright red powder (12%); mp 232.8-233.1 ° C; ¹ H-NMR (CDCl _{3 ,} 400 MHz) δ 3.91 (s, 3H, methoxy), 7.11 (d, 1H, benzene), 7.25 (m, 2H, benzene), 7.38 (t, 1H, benzene), 7.73 (d, 1H, benzene), 7.68 (q, 1H, 2-pyridine), 8.00 (s, 2H, NH 2), 8.46 (q, 1H, 2-pyridine), 9.09 (q, 1H, 2-pyridine); MS (m / z) 348 (M ⁺ ).

Compounds synthesized based on NMR analysis were identified as Methyl 2-amino-4,9-dihydro-4,9-dioxo-1-phenyl-1H-pyrrolo [3,2-g] quinoline-3-carboxylate. The synthesized compound was named "YSK2823".

Example  5: Methyl  2- (7- chloro -5,8- dihydro -5,8- dioxoisoquinolin -6- yl Synthesis of 2-cyanoacetate

A is methyl cyanoaccetate / ethanol in the above scheme.

6,7-dichloroisoquinoline-5,8-dione 590 mg (2.66 mmole) was added to 100 ml abs. After dissolving in ethanol (EtOH), 0.26358 ml (2.66 mmol) of methyl cyanoacetate was added thereto. 2 ml of ammonia water was added and reacted for 10 minutes, followed by acidification with d-HCl. Extraction with MC and separation by column chromatography (EA) with EA were performed. NMR analysis data of the synthesized compound was as follows.

Brown oil (42%); ¹ H-NMR (CDCl _3, 400 MHz) δ 3.86 (s, 3H, methoxy), 5.50 (s, 1H, CH), 7.96 (d, 1H, J = 4.8, 3-pyridine), 9.13 (d, 1H, J = 4.8, 3-pyridine ), 9.39 (s, 1 H, 3-pyridine); MS (m / z) 290 (M ⁺ ).

The synthesized compound was identified as Methyl 2- (7-chloro-5,8-dihydro-5,8-dioxoisoquinolin-6-yl) -2-cyanoacetate.

Example 6: Methyl 2-amino-4,9-dihydro-4,9-dioxo-1-p-tolyl-1H-pyrrolo [3,2-g] isoquinoline -3- carboxylate  synthesis

B in the above scheme is 4-methyl-phenylamine / ethanol.

Compound Methyl 2- (7-chloro-5,8-dihydro-5,8-dioxoisoquinolin-6-yl) -2-cyanoacetate synthesized in Example 5 150 mg 100 ml abs. After dissolving and stirring in EtOH, 4-methyl-phenylamine corresponding to the equivalent was added and reacted at room temperature for 4-5 hours. Termination of the reaction was confirmed by TCL. The reaction solution was cooled and concentrated, and then separated by column chromatography with EA: HX = 1: 1 solvent and recrystallized with 95% EtOH.

The NMR analysis of the synthesized compound was as follows.

Dark red powder (12%); mp 218.9-219.4 ° C; ¹ H-NMR (CDCl _{3 ,} 400 MHz) δ 2.48 (s, 3H, methyl), 3.96 (s, 3H, methoxy), 6.97 (m, 2H, benzene), 7.04 (m, 2H, benzene), 7.50 (s, 2H, NH2), 7.83 (d, 1H, J = 5.2, 3-pyridine), 9.07 (d, 1H, J = 5.2, 3-pyridine), 9.44 (s, 1H, 3-pyridine); MS (m / z) 361 (M ⁺ ), 332 (M ⁺ -29 -OCH ₃ ); MS (m / z) 361 (M ⁺ ).

The synthesized compound was identified as Methyl 2-amino-4,9-dihydro-4,9-dioxo-1-p-tolyl-1H-pyrrolo [3,2-g] isoquinoline-3-carboxylate and identified as “YSK2824”. Named it.

Example  7: Methyl  2- amino -1- (4- fluorophenyl ) -4,9- dihydro -4,9- dioxo -1H- pyrrolo [3,2-g] isoquinoline -3- carboxylate Synthesis of

B in the scheme is 4-fluoro-phenylamine / ethanol. Methyl 2- (7-chloro-5,8-dihydro-5,8-dioxoisoquinolin-6-yl) -2-cyanoacetate as a compound synthesized in Example 5 150 mg 100 ml abs. After dissolving in EtOH and stirring, 4-fluoro-phenylamine corresponding to the equivalent was added and reacted at room temperature for 4-5 hours. Termination of the reaction was performed using TCL, the reaction solution was cooled and concentrated, separated by column chromatography with EA: HX = 1: 1 solvent, and recrystallized with 95% EtOH.

The NMR analysis of the synthesized compound was as follows.

Dark red powder (15%); mp 241.5-241.8 ° C; ¹ H-NMR (CDCl _{3 ,} 400 MHz) δ 3.95 (s, 3H, methoxy), 7.12 (m, 4H, benzene), 7.87 (s, 2H, NH2), 7.91 (d, 1H, J = 5.2, 3-pyridine), 9.11 (d, 1H, J = 5.2, 3-pyridine), 9.45 (s, 1H, 3-pyridine); MS (m / z) 365 (M ⁺ ), 336 (M ⁺ -29 -OCH ₃ ); MS (m / z) 366 (M ⁺ ).

The synthesized compound was identified as Methyl 2-amino-4,9-dihydro-4,9-dioxo-1-p-tolyl-1H-pyrrolo [3,2-g] isoquinoline-3-carboxylate and identified as “YSK2825”. Named it.

Example  8: aorta Smooth muscle cell  Of the present invention on growth inhibition YSK2821 , YSK2822, YSK2823 , YSK2824 , YSK2825  Compound effect

4.0 x 10 ⁴ cells / ml to determine whether the indoldion derivative compounds YSK2821, YSK2822, YSK2823, YSK2824, and YSK2825 synthesized in Examples 2, 3, 4, 6, and 7 inhibited the growth of aortic smooth muscle cells in rats. Of aortic smooth muscle cells were transferred to 12 well plates and incubated for 24 hours in DMEM medium containing 0.5% (V / V) fetal calf serum. Subsequently, the medium was treated with compounds YSK2821, YSK2822, YSK2823, YSK2824, and YSK2825 at a concentration range of 0.5-5 μM, followed by further incubation for 24 hours, followed by platelet-dereived growth factor (PDGF-). BB) and after 24 hours with trypsin to separate cells from the bottom and count the number of cells separated using a hemocytometer.

Figure 1a-1e is a graph showing the results of the proliferation inhibitory effect of the compounds YSK2821, YSK2822, YSK2823, YSK2824, YSK2825 after treatment of rat aortic smooth muscle cells in the concentration range of 0.5-5 μM. From the results of FIGS. 1A-1E, it was confirmed that the indoldion derivative compounds YSK2821, YSK2822, YSK2823, YSK2824, and YSK2825 of the present invention effectively inhibit the proliferation of aortic smooth muscle cells in the concentration range of 0.5-5 μM.

Example  9: YSK2821 , YSK2822 , YSK2823 , YSK2824 , YSK2825  Aortic smooth muscle cell of compound DNA  Synthesis impact

In Example 6, the compounds YSK2821, YSK2822, YSK2823, YSK2824, YSK2825 was confirmed to inhibit the proliferation of aortic smooth muscle cells in rats. In order to confirm whether the compounds of the present invention are caused by the mechanism of inhibiting the proliferation of aortic smooth muscle cells, the effects of the compounds YSK2821, YSK2822, YSK2823, YSK2824, and YSK2825 on the cell biosynthesis were examined.

2.0 x 10 ⁴ cells / ml of aortic smooth muscle cells were transferred to 24 well plates and incubated for 24 hours in DMEM medium containing 0.5% (V / V) fetal bovine serum. Subsequently, YSK2821, YSK2822, YSK2823, YSK2824, and YSK2825 were respectively treated at concentration ranges of 0.5-5 μM, incubated for another 24 hours, and then platelet-dereived growth factor (PDGF-BB). . After 20 hours, 1 μl of [ ³ H] -thymidine was treated. After incubation for 4 hours, the plate was washed twice with PBS and treated with TCA reagent for 30 minutes. The TCA reagent was removed and washed twice with ethanol / ether (1: 1 v / v). 500 μl of 1 N NaOH was dispensed into each well to completely lyse the cells, and then mixed with a 3 ml scintillation cocktail to measure radioactivity with a liquid scintillation counter. .

Figure 2a-2e is a graph showing the effect of inhibiting the DNA synthesis of the cells after the treatment of aortic smooth muscle cells of rats in the concentration range of 0.5-5 μM YSK2821, YSK2822, YSK2823, YSK2823, YSK2824, YSK2825. From the results shown in FIGS. 2A-2E, it can be seen that the compounds YSK2821, YSK2822, YSK2823, YSK2824, and YSK2825 of the present invention effectively inhibit the biosynthesis of DNA of aortic smooth muscle cells in the concentration range of 0.5-5 μM.

Example  10: YSK2821 , YSK2822 , YSK2823 , YSK2824 , YSK2825  Effect of Compounds on Aortic Smooth Muscle Cell Proliferation Cycle

The changes in the cell cycle progression induced by treatment of the compounds YSK2821, YSK2822, YSK2823, YSK2824, and YSK2825 of the present invention on aortic smooth muscle cells induced by platelet-derived growth factor were examined. 4.0 x 10 ⁴ cells / ml of aortic smooth muscle cells were transferred to 12 well plates and incubated for 24 hours in DMEM medium containing 0.5% (V / V) fetal calf serum. Subsequently, the medium was treated with compounds YSK2821, YSK2822, YSK2823, YSK2824, and YSK2825 at a concentration range of 0.5-5 μM, followed by further incubation for 24 hours, followed by platelet-dereived growth factor (PDGF-). BB) and cultured cells were stained using PI staining. Since the amount of the nucleus was confirmed by FACS (fluorescence activated cell sorter) method.

3A-3E are graphs showing the effects of the compounds YSK2821, YSK2822, YSK2823, YSK2824, and YSK2825 on rat aortic smooth muscle cells in the concentration range of 0.5-5 μM and then on the cell proliferation cycle of these cells. From the results of FIGS. 3A-3E, the compounds YSK2821, YSK2822, YSK2823, YSK2824, and YSK2825 of the present invention retard the inter-phase of G ₀ / G ₁ and S phase in the vascular smooth muscle cell cycle progression induced by platelet-derived growth factors. It was found that the cell proliferation was suppressed.

Example  11: Acute Toxicity Study

Three groups of 25 ± 5 g ICR mice and 235 ± 10 g SPF Sprague Dawley (Biogenomics) rats were divided into four groups of YSK2821, YSK2822, YSK2823, YSK2824, and YSK2825. Toxicity was observed for 24 hours after intraperitoneal administration of 100 mg / kg, 10 mg / kg, 1 mg / kg doses respectively.

As a result of toxicity experiments, no deaths were observed in all four groups, and no symptoms were observed in weight gain and feed intake. Therefore, it was confirmed that the compound of the present invention is a safe drug.

Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that the specific technology is merely a preferred embodiment, and the scope of the present invention is not limited thereto. Thus, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

Figure 1a-1e is a graph showing the results of the proliferation inhibitory effect of the indole dione derivative compounds YSK2821, YSK2822, YSK2823, YSK2824, YSK2825 of the present invention to the aortic smooth muscle cells of rats after treatment in the concentration range of 0.5-5μM to be.

Figure 2a-2e is a graph showing the effect of inhibiting DNA synthesis of the indole dione derivative compounds YSK2821, YSK2822, YSK2823, YSK2824, YSK2825 of the present invention after treatment of rat aortic smooth muscle cells in the concentration range of 0.5-5μM .

Figure 3a-3e shows the effect of the indole dione derivative compounds YSK2821, YSK2822, YSK2823, YSK2824, YSK2825 of the present invention on the aortic smooth muscle cells of rats after concentration in the concentration of 0.5-5μM, the cell proliferation cycle of the cells It is a graph.

Claims (6)

An indoldion derivative compound represented by the following general formula (I). [Formula I]

Provided that X and Y in Formula I are each independently C or N, R ₁ when X is C and Y is N Is H or a C ₁ -C ₄ straight or branched alkyl group, R ₂ Is H, C ₁ -C ₄ linear or branched alkyl or alkoxy group, R ₁ when X is N and Y is C Is H or a C ₁ -C ₄ straight or branched alkyl group, R ₂ Is an alkyl group of H, halogen, C ₁ -C ₄ straight or branched chain. A composition for inhibiting vascular smooth muscle cell proliferation, comprising an indoldione derivative compound represented by Formula I as an active ingredient. [Formula I]

Provided that X and Y in Formula I are each independently C or N, R ₁ when X is C and Y is N Is H or a C ₁ -C ₄ straight or branched alkyl group, R ₂ is H, a C ₁ -C ₄ straight or branched alkyl or alkoxy group, When X is N and Y is C, R ₁ is H or a C ₁ -C ₄ straight or branched alkyl group, and R ₂ is H, halogen or a C ₁ -C ₄ straight or branched alkyl group. (a) a pharmaceutically effective amount of an indoldione derivative compound represented by formula (I); And (b) a pharmaceutical composition for preventing or treating vascular smooth muscle cell hyper-proliferative disorder, comprising a pharmaceutically acceptable carrier. [Formula I]

Provided that X and Y in Formula I are each independently C or N, R ₁ when X is C and Y is N Is H or a C ₁ -C ₄ straight or branched alkyl group, R ₂ is H, a C ₁ -C ₄ straight or branched alkyl or alkoxy group, When X is N and Y is C, R ₁ is H or a C ₁ -C ₄ straight or branched alkyl group, and R ₂ is H, halogen or a C ₁ -C ₄ straight or branched alkyl group. The compound of claim 3, wherein R ₁ when X is C and Y is N Is H or -CH ₃ , R ₂ Is H, -CH _{3 ,} or -OCH ₃ , and R ₁ when X is N and Y is C Is H or -CH ₃ , R ₂ Is -CH ₃ , fluorine (F), chlorine (Cl), bromine (Br) or iodine (I). The pharmaceutical composition of claim 3, wherein the compound of Formula I is any one of Formulas II to VI.  [Formula II] [Formula III] [Formula IV]

[Formula Ⅴ] [Formula VI]

The method of any one of claims 3 to 5, wherein the vascular smooth muscle cell abnormality proliferative disease includes vascular restenosis, vascular stenosis, arteriosclerosis, atherosclerosis, heart failure, myocardial infarction, angina pectoris, arrhythmia, and hypertensive heart. A pharmaceutical composition, characterized in that the disease, congenital heart disease, stroke, or peripheral vascular stenosis.

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